Correlation Between Glycolysis And Proliferationand Differentiation Of HaCaT Cells In Psoriatic Similar Inflammatory Microenvironment

ObjectiveTo investigate the status of glucose metabolism,proliferation and differentiation of HaCaT cells in psoriatic inflammatory microenvironment,and analyze the relationship between glucose metabolism,proliferation and differentiation,and further reveal the mechanism of abnormal proliferation and differentiation of KC in psoriasis.Methods1.The liquid nitrogen freezed-stored HaCaT cells were quickly put into 42℃constant temperature water bath,and resuscitated with MEM medium containing 10%fetal bovine,penicillin 100KU/L and streptomycin 100mg/L,and cultured with the same medium.After passage,when the cells were about 60% in the six-well plate.The cells were treated with M5(a mixture of IL-1α,IL-17 A,IL-22,tumor suppressant protein M and TNF-α)and PBS buffer for 48 h,and the psoriatic keratinocytes(KCs)model was simulated as the experimental group,and the normal human KCs model was the control group;2.Cell energy metabolism analyzer was used to detect the glucose metabolism level of cells in both groups,CCK-8 assay was used to detect the absorbance(OD value),RT-qPCR was used to detect the mRNA relative expression levels of K6 and K16 to indicate cell proliferation,and RT-qPCR was used to detect the mRNA relative expression levels of K1 and K10 to indicate cell differentiation;3.T test was used to analyze the differences in glucose metabolism,proliferation,and differentiation between the two groups,and Peaeson correlation analysis was used to analyze the correlation between glucose metabolism,proliferation,and differentiation.Results1.Compared with the control group,the basal glycolysis and glycolysis ability of the experimental group were significantly increased(P<0.01),while the basic OCR value and the maximum OCR value did not change significantly(P > 0.05);2.CCK8 method was used to detect cell proliferation.The results showed that OD value in the experimental group was higher than that in the control group(P<0.02);3.The relative expression levels of K6 and K16 mRNA in the experimental group were higher than those in the control group(P<0.02)4.The relative expression levels of K1 and K06 mRNA in the experimental group were higher than those in the control group(P < 0.05);5.Pearson correlation analysis showed that basal glycolysis of HaCaT cells was positively correlated with OD value,mRNA relative expression levels of K6 and K16(R value 0.679,0.757 and 0.789,P value 0.015,0.004 and 0.002).Glycolysis ability was also positively correlated with OD value,mRNA relative expression levels of K6 and K16(R value was 0.726,0.767 and 0.799,P value was 0.007,0.004 and 0.002).There was a negative correlation between basal glycolysis and the relative expression levels of K1 and K10 mRNA(R value was-0.625 and-0.606,P value was 0.030 and 0.037),and there was a negative correlation between glycolysis ability and the relative expression levels of K1 and K10 mRNA(R value was-0.662 and-0.655,P values were 0.019 and0.021).Conclusion1.The glucose metabolism pattern of psoriatic KC model of psoriasis showed significantly increased glycolysis level,excessive proliferation and abnormal differentiation.2.Abnormally elevated glycolysis level may promote KC proliferation,inhibit its differentiation,and aggravate hyperkeratosis and hypokeratosis of psoriatic epidermis through rapid production of ATP and intermediate metabolites.